LOCUS       V040009                 8927 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040009
VERSION     V040009
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..3846
                     /label="G6PD(NM_001042351)"
                     /note="G6PD(NM_001042351)"
                     /gene="G6PD"
     promoter        3897..4108
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4121..4389
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4416..5015
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5022..5610
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5684..5917
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5989..6110
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6129..6264
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6302..6318)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6326..6342
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6350..6380)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6395..6416
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6704..7292)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7463..8323)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8324..8428)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8902..8918
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catggcagag caggtggccc tgagccggac ccaggtgtgc
     2341 gggatcctgc gggaagagct tttccagggc gatgccttcc atcagtcgga tacacacata
     2401 ttcatcatca tgggtgcatc gggtgacctg gccaagaaga agatctaccc caccatctgg
     2461 tggctgttcc gggatggcct tctgcccgaa aacaccttca tcgtgggcta tgcccgttcc
     2521 cgcctcacag tggctgacat ccgcaaacag agtgagccct tcttcaaggc caccccagag
     2581 gagaagctca agctggagga cttctttgcc cgcaactcct atgtggctgg ccagtacgat
     2641 gatgcagcct cctaccagcg cctcaacagc cacatgaatg ccctccacct ggggtcacag
     2701 gccaaccgcc tcttctacct ggccttgccc ccgaccgtct acgaggccgt caccaagaac
     2761 attcacgagt cctgcatgag ccagataggc tggaaccgca tcatcgtgga gaagcccttc
     2821 gggagggacc tgcagagctc tgaccggctg tccaaccaca tctcctccct gttccgtgag
     2881 gaccagatct accgcatcga ccactacctg ggcaaggaga tggtgcagaa cctcatggtg
     2941 ctgagatttg ccaacaggat cttcggcccc atctggaacc gggacaacat cgcctgcgtt
     3001 atcctcacct tcaaggagcc ctttggcact gagggtcgcg ggggctattt cgatgaattt
     3061 gggatcatcc gggacgtgat gcagaaccac ctactgcaga tgctgtgtct ggtggccatg
     3121 gagaagcccg cctccaccaa ctcagatgac gtccgtgatg agaaggtcaa ggtgttgaaa
     3181 tgcatctcag aggtgcaggc caacaatgtg gtcctgggcc agtacgtggg gaaccccgat
     3241 ggagagggcg aggccaccaa agggtacctg gacgacccca cggtgccccg cgggtccacc
     3301 accgccactt ttgcagccgt cgtcctctat gtggagaatg agaggtggga tggggtgccc
     3361 ttcatcctgc gctgcggcaa ggccctgaac gagcgcaagg ccgaggtgag gctgcagttc
     3421 catgatgtgg ccggcgacat cttccaccag cagtgcaagc gcaacgagct ggtgatccgc
     3481 gtgcagccca acgaggccgt gtacaccaag atgatgacca agaagccggg catgttcttc
     3541 aaccccgagg agtcggagct ggacctgacc tacggcaaca gatacaagaa cgtgaagctc
     3601 cctgacgcct atgagcgcct catcctggac gtcttctgcg ggagccagat gcacttcgtg
     3661 cgcagcgacg agctccgtga ggcctggcgt attttcaccc cactgctgca ccagattgag
     3721 ctggagaagc ccaagcccat cccctatatt tatggcagcc gaggccccac ggaggcagac
     3781 gagctgatga agagagtggg tttccagtat gagggcacct acaagtgggt gaacccccac
     3841 aagctctgag gatccgcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc
     3901 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg
     3961 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     4021 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     4081 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc
     4141 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
     4201 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
     4261 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
     4321 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg
     4381 ttacagatcc aagctgtgac cggcgcctac gctagatgac cgagtacaag cccacggtgc
     4441 gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg
     4501 actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc
     4561 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     4621 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     4681 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     4741 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     4801 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     4861 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     4921 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     4981 gcacctggtg catgacccgc aagcccggtg cctgagtcga caatcaacct ctggattaca
     5041 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     5101 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     5161 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     5221 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     5281 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     5341 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     5401 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     5461 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     5521 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     5581 gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat gacttacaag
     5641 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     5701 tcccaacgaa aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     5761 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     5821 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     5881 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     5941 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     6001 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6061 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     6121 ctctagctat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6181 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     6241 agtgaggagg cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat
     6301 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6361 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6421 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6481 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6541 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6601 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6661 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6721 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6781 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6841 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6901 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6961 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7021 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7081 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7141 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7201 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7261 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7321 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7381 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7441 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7501 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7561 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7621 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7681 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7741 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7801 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7861 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7921 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7981 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8041 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     8101 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8161 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8221 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8281 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8341 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8401 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     8461 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     8521 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     8581 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     8641 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     8701 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     8761 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     8821 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     8881 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
